Targeted UHPLC-ESI-MS/MS Analysis of Selected Neurotransmitters, Tryptophan and Its Metabolite Kynurenine in Tau Transgenic Rat Brain Tissue: A Pivotal Study

Neurotransmitters (NT) are widely distributed in the central nervous system. These molecules are important for many physiological processes and the function of the immune system. Imbalance of NT are linked to numerous neurological disorders and diseases, including tauopathies. Here, a targeted approach based on on-line combination of ultra-high performance liquid chromatography with tandem mass spectrometry was validated and applied to the quantitative analysis of nine NT (acetylcholine, choline, aspartic acid, asparagine, glutamic acid, glutamine, pyroglutamate, γ-aminobutyric acid, N-acetyl-L-aspartic acid), tryptophan and its metabolite kynurenine in brain tissue samples of a rat model for tauopathy. The applied analytical method was characterized by excellent validation parameters for all analytes, such as limits of detection in the range of 0.01–1.70 µg/mL, regression coefficients of the calibration curves ≥ 0.9946, intra-day and inter-day precision expressed as coefficient of variation in the range of 0.6–11.9% and 0.6–14.4%, and accuracy in the range of 87.6–107.1% and 87.2–119.6%. Our analytical approach led to the identification of increased levels of choline and γ-aminobutyric acid in pons, and elevated concentration levels of pyroglutamate in medulla oblongata. These findings indicate that NT could play a valuable role in the study and clarification of neuroinflammation and neurodegenerative diseases.

Augmentation of extracellular ATP synergizes with chemotherapy in triple negative breast cancer

Introduction: Breast cancer affects two million women worldwide every year and is the most common cause of cancer-related death among women. The triple-negative breast cancer (TNBC) sub-type is associated with an especially poor prognosis because currently available therapies, fail to induce long-lasting responses. Therefore, there is an urgent need to develop novel therapies that result in durable responses. One universal characteristic of the tumor microenvironment is a markedly elevated concentration of extracellular adenosine triphosphate (eATP). Chemotherapy exposure results in further increases in eATP through its release into the extracellular space of cancer cells via P2RX channels. eATP levels are reduced by eATPases. Given that high concentrations of eATP are cytotoxic, we hypothesized that augmenting the release of eATP through P2RX channels and inhibiting extracellular ATPases would sensitize TNBC cells to chemotherapy. Methods: TNBC cell lines MDA-MB 231, Hs 578t and MDA-MB 468 and non-tumorigenic immortalized mammary epithelial MCF-10A cells were treated with increasing concentrations the chemotherapeutic agent paclitaxel in the presence of eATPase inhibitors, specific agonists or antagonists of P2RXs with cell viability and eATP content being measured. Additionally, the mRNA, protein and cell surface expressions of the purinergic receptors P2RX4 and P2RX7 were evaluated in all examined cell lines via qRT-PCR, western blot, and flow cytometry analyses, respectively. Results: In the present study, we observed dose-dependent declines in cell viability and increases in eATP in paclitaxel-treated TNBC cell lines in the presence of inhibitors of eATPases. These effects were reversed by specific antagonists of P2RXs. Similar results were observed with P2RX activators. All examined cell lines expressed both P2RX4 and P2RX7 at the mRNA, protein and cell surface levels. Conclusion: These results reveal that eATP modulates the chemotherapeutic response in TNBC cell lines which could be exploited to enhance the efficacy of chemotherapy regimens for TNBC.

Interplay of Dinner Timing and MTNR1B Type 2 Diabetes Risk Variant on Glucose Tolerance and Insulin Secretion: A Randomized Crossover Trial

<strong>Objective: </strong>We tested whether the concurrence of food intake and elevated concentration of endogenous melatonin, as occurs in late eating, results in impaired glucose control, in particular in carriers of the type 2 diabetes-associated G allele in the melatonin-receptor-1-b gene (<i>MTNR1B</i>).<strong> </strong> <p><strong>Research Design and Methods:</strong> In a Spanish natural late eating population, a randomized, cross-over study design was performed, following an 8-h fast. Each participant <strong>(n=845) </strong>underwent two evening 2-h 75g oral glucose tolerance tests (OGTT): an early condition scheduled 4 hours prior to habitual bedtime <strong>(“early dinner-timing”)</strong>, and a late condition scheduled 1 hour prior to habitual bedtime <strong>(“late dinner-timing”)</strong>, simulating an early and a late dinner timing, respectively.<strong> </strong>Differences in postprandial glucose and insulin responses were determined using incremental area under the curve (AUC) calculated by the trapezoidal method between <strong>early and late dinner-timing.</strong><strong></strong></p> <p><strong>Results:</strong> <strong>Melatonin serum levels were </strong>3.5-fold <strong>higher in the late <i>vs. </i>early condition, with late dinner-timing resulting in </strong>6.7% <strong>lower insulin</strong> <strong>area-under-the-curve (AUC) and </strong>8.3%<strong> higher glucose</strong> <strong>AUC. In the late condition<i> MTNR1B</i> G-allele carriers had lower glucose tolerance than non-carriers. Genotype differences in glucose tolerance were attributed to reductions in </strong>β-cell <strong>function (<i>P_int</i>AUCgluc=0.009, <i>P_int</i>CIR=0.022, <i>P_int</i>DI=0.018).</strong></p> <p><strong>Conclusions:</strong> <strong>Concurrently high endogenous melatonin and carbohydrate intake, as typical for late eating, impair glucose tolerance, especially in <i>MTNR1B</i> G-risk-allele carriers<i>, </i>attributable to insulin secretion defects.</strong></p>

Interplay of Dinner Timing and MTNR1B Type 2 Diabetes Risk Variant on Glucose Tolerance and Insulin Secretion: A Randomized Crossover Trial

<strong>Objective: </strong>We tested whether the concurrence of food intake and elevated concentration of endogenous melatonin, as occurs in late eating, results in impaired glucose control, in particular in carriers of the type 2 diabetes-associated G allele in the melatonin-receptor-1-b gene (<i>MTNR1B</i>).<strong> </strong> <p><strong>Research Design and Methods:</strong> In a Spanish natural late eating population, a randomized, cross-over study design was performed, following an 8-h fast. Each participant <strong>(n=845) </strong>underwent two evening 2-h 75g oral glucose tolerance tests (OGTT): an early condition scheduled 4 hours prior to habitual bedtime <strong>(“early dinner-timing”)</strong>, and a late condition scheduled 1 hour prior to habitual bedtime <strong>(“late dinner-timing”)</strong>, simulating an early and a late dinner timing, respectively.<strong> </strong>Differences in postprandial glucose and insulin responses were determined using incremental area under the curve (AUC) calculated by the trapezoidal method between <strong>early and late dinner-timing.</strong><strong></strong></p> <p><strong>Results:</strong> <strong>Melatonin serum levels were </strong>3.5-fold <strong>higher in the late <i>vs. </i>early condition, with late dinner-timing resulting in </strong>6.7% <strong>lower insulin</strong> <strong>area-under-the-curve (AUC) and </strong>8.3%<strong> higher glucose</strong> <strong>AUC. In the late condition<i> MTNR1B</i> G-allele carriers had lower glucose tolerance than non-carriers. Genotype differences in glucose tolerance were attributed to reductions in </strong>β-cell <strong>function (<i>P_int</i>AUCgluc=0.009, <i>P_int</i>CIR=0.022, <i>P_int</i>DI=0.018).</strong></p> <p><strong>Conclusions:</strong> <strong>Concurrently high endogenous melatonin and carbohydrate intake, as typical for late eating, impair glucose tolerance, especially in <i>MTNR1B</i> G-risk-allele carriers<i>, </i>attributable to insulin secretion defects.</strong></p>

Interplay of Dinner Timing and MTNR1B Type 2 Diabetes Risk Variant on Glucose Tolerance and Insulin Secretion: A Randomized Crossover Trial

OBJECTIVE We tested whether the concurrence of food intake and elevated concentration of endogenous melatonin, as occurs in late eating, results in impaired glucose control, in particular in carriers of the type 2 diabetes–associated G allele in the melatonin receptor-1b gene (MTNR1B). RESEARCH DESIGN AND METHODS In a Spanish natural late-eating population, a randomized, crossover study was performed. Each participant (n = 845) underwent two evening 2-h 75-g oral glucose tolerance tests following an 8-h fast: an early condition scheduled 4 h prior to habitual bedtime (“early dinner timing”) and a late condition scheduled 1 h prior to habitual bedtime (“late dinner timing”), simulating an early and a late dinner timing, respectively. Differences in postprandial glucose and insulin responsesbetween early and late dinner timing were determined using incremental area under the curve (AUC) calculated by the trapezoidal method. RESULTS Melatonin serum levels were 3.5-fold higher in the late versus early condition, with late dinner timing resulting in 6.7% lower insulin AUC and 8.3% higher glucose AUC. In the late condition, MTNR1B G-allele carriers had lower glucose tolerance than noncarriers. Genotype differences in glucose tolerance were attributed to reductions in β-cell function (P for interaction, Pint glucose area under the curve = 0.009, Pint corrected insulin response = 0.022, and Pint Disposition Index = 0.018). CONCLUSIONS Concurrently high endogenous melatonin and carbohydrate intake, as typical for late eating, impairs glucose tolerance, especially in MTNR1B G-risk allele carriers, attributable to insulin secretion defects.

Extract of Trema Orientalis (L.) Stem Bark: A Potential Source of Environmentally Friendly Tanning Agent for Leather Industry

Researchers around the world are in continuous endeavor to develop environment friendly tanning agent due to adverse effect of conventionally used chromium during leather tanning. Recent trend of tanning is thus heading to chrome-free and greener chemical processing options. Vegetable tanning is an exoteric leather processing technique because of their lower pollution load on the environment. Considering the inadequacy and high costing for the commercialized tannins, development of alternative tannins from locally available plants and their characterizations are important. In this research, the stem barks of Trema Orientalis (L.) were extracted at different temperatures employing water solvent with or without additives (sodium hydroxide or sodium sulphite) to attain phenolic-rich extractives. To ascertain appropriateness as a vegetable tanning agent, the obtained extracts were thereafter characterized in respect to yield, total phenolic content, tannin content and molecular structure. The extraction yield for all extracts improves and the quality remains nearly unchanged with temperature rising. The elevated concentration of chemical additives enhances the extraction yield but lessens the quality of extracts. In terms of extraction yield and the quality of extracts the best condition for extraction was discerned at 80°C with water solvent. This water extract has a decent extent of phenolic and tannin content of 266.13 mg Gallic acid equivalent/ gm of dry extract and 30.12 % respectively. The final extract exhibits excellent leather retanning tendency comparable to the commercial quebracho tannins.

Effectiveness of Zr(IV)-Loaded Banana Peels Biomass for the Uptake of Fluoride Anion from Water

The present study reports the fluoride uptake potential of Zr(IV)-loaded saponified banana peels (Zr(IV)-SBP) from water. Zr(IV)-SBP was synthesized by loading Zr(IV) onto banana peel biomass after saponification and sorbent characterization was performed by using different techniques including FE-SEM (Field Emission Scanning Electron Microscopy), FTIR (Fourier Transform Infra-Red) spectroscopy and zeta potential analysis. Batch experiments were carried out to examine the monitoring factors for the uptake of fluoride onto the investigated adsorbent. The optimal pH and contact time were found to be 2.94 and 300 minutes, respectively. The results from characterization techniques concurred that Zr(IV)-SBP have prominent adsorption sites favorable for the sorption of fluoride ions. The sorption behavior of fluoride onto Zr(IV)-SBP was best fitted with the Langmuir adsorption isotherm and pseudo-second-order kinetics model. The maximum adsorption capacity of Zr(IV)-SBP was 36.02 mg/g using the Langmuir isotherm model. The coexisting ions like chloride and nitrate caused very small interference, elevated concentration of sulphate notably lowers the fluoride adsorption percentage in the binary system, and the sorption using multiple systems was lowered significantly which is due to the synergistic effect of co-existing interfering ion. The adsorbed fluoride was completely desorbed using 2M NaOH solution. Fluoride sorption performance of Zr(IV)-SBP demonstrated that it can be a low cost, environmentally benign and one of the highly potent alternatives for the remediation of fluoride ions to avoid ablation on the water.

Determination of Floral Origin Markers of Latvian Honey by Using IRMS, UHPLC-HRMS, and 1H-NMR

The economic significance of honey production is crucial; therefore, modern and efficient methods of authentication are needed. During the last decade, various data processing methods and a combination of several instrumental methods have been increasingly used in food analysis. In this study, the chemical composition of monofloral buckwheat (Fagopyrum esculentum), clover (Trifolium repens), heather (Calluna vulgaris), linden (Tilia cordata), rapeseed (Brassica napus), willow (Salix cinerea), and polyfloral honey samples of Latvian origin were investigated using several instrumental analysis methods. The data from light stable isotope ratio mass spectrometry (IRMS), ultra-high performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS), and nuclear magnetic resonance (NMR) analysis methods were used in combination with multivariate analysis to characterize honey samples originating from Latvia. Results were processed using the principal component analysis (PCA) to study the potential possibilities of evaluating the differences between honey of different floral origins. The results indicate the possibility of strong differentiation of heather and buckwheat honeys, and minor differentiation of linden honey from polyfloral honey types. The main indicators include depleted δ15N values for heather honey protein, elevated concentration levels of rutin for buckwheat honey, and qualitative presence of specific biomarkers within NMR for linden honey.

Assessing Health Risk Associated with Uranium in Rietspruit Water, Far West Rand Goldfield, South Africa

Uranium, U, as a toxic element has detrimental radiological and chemical impacts on human health when ingested at elevated concentration in water. The establishment of gold mining operations upstream to the Rietspruit is assumed to be a source of uranium into this water body which flows through residential and agricultural land. Water samples from the stream were collected and analysed using inductively coupled plasma mass spectrometry to determine the concentration of U. Uranium concentrations above South African regulatory limit were recorded in the stream water which is used for domestic, agricultural and religious activities as observed during field visits. Results from calculations showed that the maximum annual effective dose within this mining environment was above 1 mSv/year for all age groups. Furthermore, the probability of radiological risk is lower than the chemical risk which is due to the ingestion of U in the Rietspruit water. The chemical risk analysis showed that within 24 km of this mining environment, stream users are in danger of U chemical toxins therefore, it is important to educate the stream users about their health risk of U and recommend adequate interventions by relevant government agencies to this effect.

Toxicity of meta-Tyrosine

L-Tyrosine (Tyr) is one of the twenty proteinogenic amino acids and also acts as a precursor for secondary metabolites. Tyr is prone to modifications, especially under conditions of cellular redox imbalance. The oxidation of Tyr precursor phenylalanine leads to the formation of Tyr non-proteinogenic isomers, including meta-Tyr (m-Tyr), a marker of oxidative stress. The aim of this review is to summarize the current knowledge on m-Tyr toxicity. The direct m-Tyr mode of action is linked to its incorporation into proteins, resulting in their improper conformation. Furthermore, m-Tyr produced by some plants as an allelochemical impacts the growth and development of neighboring organisms. In plants, the direct harmful effect of m-Tyr is due to its modification of the proteins structure, whereas its indirect action is linked to the disruption of reactive oxygen and nitrogen species metabolism. In humans, the elevated concentration of m-Tyr is characteristic of various diseases and ageing. Indeed, m-Tyr is believed to play an important role in cancer physiology. Thus, since, in animal cells, m-Tyr is formed directly in response to oxidative stress, whereas, in plants, m-Tyr is also synthesized enzymatically and serves as a chemical weapon in plant–plant competition, the general concept of m-Tyr role in living organisms should be specified.